Open AccessEvaluation of a p30 Gene‐Based Real‐Time Reverse Transcriptase Polymerase Chain Reaction Assay for Detection of Feline Caliciviruses
Author(s) -
Scansen Brian A.,
Wise Annabel G.,
Kruger John M.,
Venta Patrick J.,
Maes Roger K.
Publication year - 2004
Publication title -
journal of veterinary internal medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.356
H-Index - 103
eISSN - 1939-1676
pISSN - 0891-6640
DOI - 10.1111/j.1939-1676.2004.tb00150.x
Subject(s) - feline calicivirus , calicivirus , polymerase chain reaction , reverse transcriptase , caliciviridae , virology , microbiology and biotechnology , sybr green i , real time polymerase chain reaction , gene , reverse transcription polymerase chain reaction , biology , titer , primer (cosmetics) , virus , gene expression , chemistry , genetics , norovirus , organic chemistry
This report describes a feline calicivirus (FCV) p30 gene‐based real‐time SYBR Green I reverse transcriptase polymerase chain reaction (RT‐PCR) assay that is capable of detecting low virus concentrations and a broad range of FCV isolates. The assay consisted of a 1‐step RT‐PCR reaction with primers delineating a 126‐base‐pair (bp) region of the FCV p30 gene. Sensitivity of the RT‐PCR assay was determined to be equivalent to a FCV titer of 1.2 × 10 1 to 1.2 × 10 2 TCID 50 /mL. The assay was linear over a wide range of template concentrations and had a reaction efficiency of 95%. Specific FCV amplification products were detected from 51 wild‐type FCV isolates, whereas specific products were not detected from a canine calicivirus, a rabbit calicivirus, and a bovine calicivirus. The primers used in this study amplified a large number of North American FCV isolates and further confirm the diagnostic utility of p30 gene‐based real‐time RT‐PCR for detection of FCV.